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UNIVERSITI PUTRA MALAYSIA PERFORMANCE OF Trichoderma harzianum Rifai AS A BIOLOGICAL CONTROL AGENT FOR BASAL STEM ROT OF OIL PALM (Elaeis guineensis Jacq.) CAUSED BY Ganoderma boninense Pat. SHAMALA A/P SUNDRAM FPSK(M) 2005 6

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UNIVERSITI PUTRA MALAYSIA

PERFORMANCE OF Trichoderma harzianum Rifai AS A BIOLOGICAL CONTROL AGENT FOR BASAL STEM ROT OF OIL PALM (Elaeis

guineensis Jacq.) CAUSED BY Ganoderma boninense Pat.

SHAMALA A/P SUNDRAM

FPSK(M) 2005 6

PERFORMANCE OF Trichoderma harzianum Rifai AS A BIOLOGICAL CONTROL AGENT FOR BASAL STEM ROT OF OIL PALM (Elaeis guineensis

Jacq.) CAUSED BY Ganoderma boninense Pat.

SHAMALA AIP SUNDRAM

Thesis Submitted to the School of Graduate Studies, Universiti Putra Malaysia in Fulfilment of the Requirements for the Degree of Master of Science

February 2005

This thesis is especially dedicated to my fam ily . . . . . ....

Abstract of thesis presented to the Senate of Universiti Putra Malaysia in fulfilment of the requirement for the degree of Master of Science

PERFORMANCE OF Trichoderma harzianum Rifai AS A BIOLOGICAL CONTROL AGENT FOR BASAL STEM ROT OF OIL PALM (Elaeis guineensis

Jacq.) CAUSED BY Ganoderma boninense Pat.

SHAMALA SUNDRAM

February 2005

Chairman : Associate Professor Faridah Abdullah, PhD

Faculty : Science

Basal stem rot (BSR) is a major threat to the oil palm industry. The disease is caused by

Ganoderma boninense, which rots the internal tissues at the trunk base resulting in stem

fracture and death of palm. The present study investigated the efficacy of two isolates of

Trichoderma harzianum (BIO T32 and BIO T66) as potential biological control agents

against BSR based on in vitro and in vivo trials.

The study revealed that treatment applied as a soil drench using conidial suspension

(mean of 1.61 x lo8 sporeslml) of BIO T32 in addition to a Trichoderma-incorporated

palm press fibre (ppf) surface mulch, performed better with a significant difference

compared to the use of BIO T66. The disease severity index (DSI) of the former was

28.35 compared to 76.67 of the latter. BIO T32 was also a competent biological control

agent in the delayed treatment given to pre-infected seedlings at 6 weeks before treatment

with BIO T32, giving a DSI of 45, which was statistically significant compared to the

infected and untreated control seedlings with a DSI of 86.67. In testing the synergistic

effect by combining the 2 isolates, a poorer performance was observed based on the DSI

and plant biomass compared to single application of BIO T32. Isolate BIO T66 which

showed good antagonistic properties in the in vitro assessment was not found to display

similar results in the in vivo trials.

A series of treatments were evaluated for their potential as a BIO T32 carrier. Out of the

3 studies, only ppf and compost exhibited promising results in their capacity as surface

4

mulches, where treatments with either one gave a DSI of 30. Both are food base carriers

for they increased the growth of oil palm seedlings significantly, with compost displaying

better results. Treatment with compost in terms of vegetative growth gave the highest

plant biomass, leaf area measurement, nitrogen, phosphorus and potassium (NPK)

content in the seedlings compared to the other 2 treatments of ppf and the untreated

control seedlings.

1

In total, the experiment revealed that the application of BIO T32 as a single inoculum

was the best treatment, giving a DSI of 28.35. Trials using a single application of BIO

T66 and BIO T66 mixed with BIO T32 performed poorly, giving a DSI of 76.67 each and

were not significantly different from the infected non-treated control plants. An

appropriate interval of conidial suspension's application played a pertinent role in the

inhibition of disease as demonstrated in the delayed treatment. The application of

compost was found to be an interesting alternative to ppf as surface mulch, which

functions also as a Trichoderma carrier. Finally, in terms of vegetative growth both ppf

and compost as food base carriers significantly increased plant biomass, total leaf area

measurement and N uptake compared to the untreated control.

Abstrak tesis yang dikemukakan kepada Senat Universiti Putra Malaysia sebagai memenuhi keperluan untuk ijazah Master Sains

KEBOLEHAN Trichoderma harzianum Rifai SEBAGAI AGEN KAWAL AN BIOLOGI TERHADAP PENYAKIT REPUT PANGKAL BATANG KELAPA

SAWIT (Elaeis guineensis Jacq.) YANG DISEBABKAN OLEH Ganoderma boninense Pat.

Oleh

SHAMALA SUNDRAM

Februari 2005

Pengerusi : Profesor Madya Faridah Abdullah, PhD

Fakulti : Sains

Penyakit reput pangkal batang adalah serius bagi industri pokok kelapa sawit. Penyakit

ini disebabkan oleh Ganoderma boninense yang menyebabkan reputan pada pangkal

pokok yang mengakibatkan kerosakan batang pokok dan akhirnya kematian. Kajian ini

menyiasat 2 isolat Trichodevma harzianum (BIO T32 dan BIO T66) dalam kebolehan

sebagai agen kawalan biologi yang berpotensi terhadap penyakit reput pangkal melalui

ujian makmal dan rurnah hijau..

Kajian ini telah membuktikan rawatan aplikasi tunggal menggunakan suspensi konidia

(purata 1.61 x lo8 spora/ml) bersama serabut kelapa sawit BIO T32 memberikan

keputusan yang lebih baik berbanding dengan BIO T66. Keputusan keparahan index

penyakit (DSI) bagi BIO T32 adalah lebih baik dengan 28.35 berbanding BIO T66

dengan 76.67. BIO T32 juga terbukti sebagai agen kawalan biologi yang berpotensi

apabila ia memberikan keputusan yang memberangsangkan bagi rawatan 6 minggu lewat

kepada anak pokok kelapa sawit yang dijangkiti EGB 01 berbanding dengan anak pokok

yang tidak dirawat (Kawalan 11). Dalam menentukan keberkesanan kombinasi 2 isolate

tersebut keputusan yang tidak baik diperolehi melalui DSI, berat kering anak pokok jika

dibandingkan dengan rawatan menggunakan BIO T32. Isolat BIO T66 yang memberikan

keputusan baik dalarn ujian in vitro tidak memberikan keputusan yang sama dalam ujian

in vivo.

4

Beberapa rawatan telah dikaji sebagai pengangkut BIO T32. Dari 3 rawatan yang dikaji,

hanya serabut kelapa sawit dan kompos memberikan keputusan yang memberangsangkan

dengan DSI 30 untuk kedua-dua rawatan di akhir eksperimen. Kedua-dua agen

pengangkut ini juga telah meningkatan pertumbuhan anak pokok dengan kompos

memberikan keputusan yang lebih tinggi dan bererti dalam penyerapan nitrogen, fosforus

dan kalium (NPK), berat kering anak pokok serta jumlah luas daun yang paling tinggi

berbanding dengan 2 rawatan iaitu serabut kelapa sawit dan kawalan (tidak dirawat).

Secara keseluruhan, kajian ini mendapati rawatan menggunakan BIO T32 secara tunggal

lebih berkesan dengan DSI 28.35. Rawatan menggunakan BIO T66 dan kombinasi

bersama BIO T32 tidak memberikan keputusan yang memberangsangkan dengan DSI

bernilai 76.67 untuk setiap satu. Suspensi konidia yang diberikan pada masa yang betul

memberikan keputusan yang memberangsangkan dalam supresi penyakit terutamanya

pada anak pokok yang dirawat lewat setelah dijangkiti awal oleh G. boninense. Kompos

menunjukkan keputusan yang memberangsangkan sebagai agen pengangkut altematif

kepada serabut kelapa sawit. Akhir sekali, kedua-dua agen iaitu serabut kelapa sawit dan

kompos sebagai pengangkut memberikan keputusan yang bererti dalarn meningkatkan

berat kering anak pokok, jumlah has daun dan pengambilan nitrogen (N), jika

dibandingkan dengan anak pokok yang tidak dirawat.

ACKNOWLEDGEMENTS

I wish to express my heartfelt thanks to my Masters Program supervisor Associate

Professor Dr. Faridah Abdullah for her constant, continuous and invaluable advice,

motivation and encouragement throughout the course of this study. I am most indebted

for her invaluable information and her constant guidance towards the completion of this

thesis. A thank you note also goes to Associate Professor Dr Umi Kalsom Yusof and

Associate Professor Dr Zainal Abidin for their support.

I would also like to express my sincere appreciation to Associate Professor Dr. Vijaya

Kanapathipillai and Dr. G. M. N. Illias for their support and encouragement in the

completion of this thesis. A thank you note goes to my labmate, Miss Jayanthi Nagappan.

My invaluable gratitude goes to my family especially to my mother for her endless effort

in persuading me to complete this thesis and not forgetting my sister Subha and brothers b

Sharma and Vignes for their support and encouragement throughout the finishing point of

this thesis. A special thank you goes to Ms Cristina Banjamin for her help and support.

Finally, a special heartfelt appreciation goes to my husband Prajiv, for his endless

motivation, assistance, continuous encouragement and guidance during the process of

completing this thesis. Thank you for everything.

I certify that an Examination Committee met on 2 " February 2005 to conduct the final examination of Sharnala Sundram on her Master of Science thesis entitled "Performance of Trichoderma harzianum Rifai as a Biological Control Agent for Basal Stem Rot of Oil Palm (Elaeis guineensis Jacq.) Caused by Ganoderma boninense Pat." in accordance with Universiti Pertanian Malaysia (Higher Degree) Act 1980 and Universiti Pertanian Malaysia (Higher Degree) Regulations 198 1. The Committee recommends that the candidate be awarded the relevant degree. Members of the Examination Committee are as follows:

Jambari Hj Ali, PhD Associate Professor Faculty of Science Universiti Putra Malaysia (Chairman)

Muskhazli Mustafa, PhD Faculty of Science Universiti Putra Malaysia (Internal Examiner)

Jugah Kadir, PhD Associate Professor Faculty of Agriculture Universiti Putra Malaysia (Internal Examiner)

Vikineswary, PhD Professor Faculty of Science Universiti Malaya (External Examiner)

Universiti Putra Malaysia

Date: 19 MAY 2005

This thesis submitted to the Senate of Universiti Putra Malaysia and has been accepted as fulfillment of the requirement for the degree of Doctor of Philosophy. The members of the Supervisory Committee are as follows:

Faridah Abdullah, PhD Associate Professor Faculty of Science Universiti Putra Malaysia (Chairman)

Umi Kalsom Yusof, PhD Associate Professor Faculty of Science Universiti Putra Malaysia (Member)

Zainal Abidin Mior Ahmad, PhD Associate Professor Faculty of Agriculture Universiti Putra Malaysia (Member)

Aini Ideris, Ph.D. Professor/Dean School of Graduate Studies

Date: 0 9 3UN 7~

DECLARATION

I hereby declare that the thesis is based on my original work except for quotations and citations which have been duly acknowledged. I also declare that it has not been previously or concurrently submitted for any other degree at UPM or other institutions

SHAMALA SUNDRAM

Date: ld t lnAY ~ 0 s

TABLE OF CONTENTS

Page

DEDICATION ABSTRACT ABSTRAK ACKNOWLEDGEMENTS APPROVAL DECLARATION LIST OF TABLES LIST OF FIGURES LIST OF ABBREVATIONS/NOTATIONS/GLOSSARY OF TERMS

CHAPTER

INTRODUCTION

LITERATURE REVIEW The Oil Palm (Elaeis guineensis Jacq.)

Origin Oil Palm Industry in Malaysia

The Botany of Oil Palm Oil Palm Environment

Soil Type Nutrients Climate

Oil Palm Diseases Root and Butt Rots Stem Diseases Leaf Diseases Diseases of the Fruit and Inflorescences Basal Stem Rot (BSR)

Predisposition Factors On The BSR Disease Age of Palms Previous Crop Types of Soils Nutrients in Soil Techniques of Replanting

Biological Control Trichoderma - Taxonomy and Morphology Trichoderma as Biological Control Agent - Previous Work Occurrence and Distribution of Trichodevma

v1 ix X

xii xvii xviii xxiii

Endurance and Propagation of Trichoderma in Soil and Plant Rhizosphere Mechanism Involved in Trichoderma Antagonism

MORPHOLOGICAL CHARACTERISTICS, ANTAGONISTIC STUDIES AND PRODUCTION OF DIFFUSIBLE METABOLITES BY SELECTED TRICHODERMA ISOLATES Introduction Materials and Methods

Source of Trichoderma Isolates Colony Characteristics and Culture Morphology Slide Cultures for Microscopic Characteristics Effect of Varying Temperature on Trichoderma Isolates Radial Growth Effect of Varying pH on Trichoderma Isolates Radial Growth Effect of Varying pH on Trichoderma Isolates Sporulation Antagonistic Test by Dual Culture Bilayer Plate Technique to Test Production of Diffusible Metabolites

Results Colony Morphology and Microscopic Characteristics from Slide Culture Trichoderma harzianum (Rifai) - BIO T32 Trichoderma harzianum (Rifai) - BIO T66 Trichoderma longibrachiatum (Rifai) - BIO T28 Trichoderma virens (Miller, Giddens & Foster) - BIO T128 4

Effect of Varying Temperature on Trichoderma Isolates Radial Growth Effect of Varying pH on Trichoderma Isolates Radial Growth Effect of Varying pH on Trichoderma Isolates Sporulation Antagonistic Test by Dual Culture Bilayer Plate Technique: Production of Diffusible Metabolites

Discussion

THE USE OF TRICHODERMA HARZIANUM AS A SINGLE 66 AND A MIXED INOCULUM SOURCE FOR THE SUPPRESSION OF BSR IN GREENHOUSE TRIALS Introduction Materials and Methods

Source of Fungal Culture Source of Seedlings Source of Potting Media Preparation of Woodblock Inocula of G, boninense (EGB 01) Preparation of Trichoderma - incorporated surface mulch Preparation of Trichoderma Conidial Suspension Artificial Infection on Oil Palm Seedlings Experimental Layout Assessment of Disease Development Disease Severity Index (DSI) Dry Weight of Plants Estimation of Spore Counts Environmental Factors: Soil Moisture Content and pH Statistical Analysis

Results Progression of Signs & Symptoms Corresponding to the Disease Classes Disease Progression Based on Disease Severity Index P S I ) 8 6 Dry Weight of Plants 88 Estimation of Trichoderma Spores: Colony Forming Unit Per Gram Soil 93 Soil Moisture and pH 9 5

Discussion 9 8

SELECTION OF AN EFFECTIVE DELIVERY SYSTEM FOR 105 T. HARZIANUM (BIO T32) AND EFFECT OF 2 DELIVERY CARRIERS ON GROWTH OF OIb PALM SEEDLINGS Introduction Materials and Methods

Source of Fungal Cultures, Oil Palm Seedlings and Potting Media Preparation of Woodblocks Inocula of EGB 01 and Method of Infection Preparation of Trichoderma - Incorporated Surface Mulch 109 Preparation of Trichoderma - Incorporated Compost 109 Preparation of Trichoderma Conidial Suspension 110 Preparation of Hydrogel Suspension 110 Experiment Layout - Effective Delivery System of BIO T32 111 Assessment of Effective Delivery System for BIO T32 11 3 Disease Severity Index (DSI) 113 Dry Weight of Plants 113

Estimation of Trichoderma Spore Count Environmental Factors: Soil Moisture and pH Statitical Analysis Experimental layout - Effect of Compost and Surface Mulch on the Growth of Oil Palm Seedlings Assessment of Oil Palm Seedlings Growth Leaf Area Measurement Dry Weight of Plants Nitrogen, Phosphorus and Potassium (NPK) Content in Leaves Statistical Analysis

Results Effective Delivery System for T. harzianum (BIO T32) Against

EGB 01 Disease Establishment based on Progression of Disease Severity Index (DSI) Effective Delivery System: Plant Dry Weight Estimation of Trichoderma Spores by cWg soil Soil Moisture and pH

Effect of Surface Mulch and Compost on Plant Growth Dry Weight of Plants Leaf Area Measurement Nitrogen, Phosphorus and Potassium (NPK) Content

Discussion

GENERAL DISCUSSION AND CONCLUSION

REFERENCES APPENDICES BIODATA OF THE AUTHOR

LIST OF TABLES

Table Page

3.1 Concentrations of lactic acid and NaoH (250ml for each pH)

3.2 Volume of Benlate @ added in PDA for the respective concentration 40

3.3 Index score for growth of EGB 01 - Bilayer Plate Technique 41 (Etheridge and Craig, 1973)

3.4 Effect of varying temperatures on Trichoderma isolates radial 51 growth (pH: 5.68)

3.5 Effect of varying pH on Trichoderma isolates radial growth 53 (temperature: 28k°C)

3.6 Effect of varying pH on Trichoderma isolates on sporulation 54 (temperature: 28*"C)

3.7 Percentage of radial inhibition (PIRG) and colony overgrowth by 58 Trichoderma test isolates

3.8 Mean summary on growth index of (EGB 01) on bilayer plates

4.1 Disease signs and symptoms corresponding to the disease class

4.2 Sequence of disease establishment on Control I1 seedlings 85 according to disease class

Mean of soil moisture percentage (%) for each group over 22 w. a. i, at 5 and 15 cm depth respectively

Mean of pH for each group over 22 w. a. i, respectively at 5 and 15 cm depth respectively

Progression of Disease Severity Index (DSI) after 24 w. a. i

Mean of soil moisture percentage (%) for each group over 22 w.a. i.

Mean reading of pH for each group over 22 w. a. i, for 5 and 15 cm depth respectively

LIST OF FIGURES

Figure

3.1 A diagram of moist chamber holding slide culture of Trichoderma isolates

Page

3 4

3.2 An illustration of measurement of radii R1 (top) and (R2) 3 8 (bottom) of G. boninense (EGB 01) used in the calculation of PIRG

3.3 Diagrammatic representation of bilayer plate technique to detect the production and effect of diffusible metabolites from Trichoderma Isolates

3.4 Surface and undersurface characteristics of BIO T32 on PDA (A) and microscopic characteristics of BIO T32 showing conidia (B) and conidiophores (C)

3.5 Surface and undersurface characteristics of BIO T66 on PDA (A) and microscopic characteristics of BIO T66 showing conidia (B) and conidiophores (C)

3.6 Surface and undersurface characteristics of BIO T28 on PDA 48 (A) and microscopic characteristics of BIO T28 showing conidia (B) and conidiophores (C)

3.7 Surface and undersurface characteristics of BIO T128 on PDA 49 (A) and microscopic characteristics of BIO TI28 showing conidia (B) and conidiophores (C) a

3.8 Surface and Undersurface of test pathogen EGB 01 (G. boninense)

3.9 Effect of temperature on radial growth of Trichoderma isolates A: BIO T32, B: BIO T66, C: BIO T28 and D: BIO T128. Plates arranged from left to right: 15"C, 25"C, 28°C (middle plate), 35°C and 40°C

3.10 Effect of varying pH on radial growth and sporulation of BIO 55 T32. Top from left to right: 2.7, 3.0,4.0, 5.0; Middle: 5.68 - Control and Bottom; from left to right: 6.0,7.0,7.6 and 8.0

xviii

r-7

3.1 1 Effect of varymg pH on radial growth and sporulation of BIO T66. Top from left to right: 2.7, 3.0,4.0, 5.0; Middle: 5.68 - Control and Bottom; from left to right: 6.0,7.0,7.6 and 8.0

3.12 Effect of varying pH on radial growth and sporulation of BIO T28. Top from left to right: 2.7, 3.0,4.0, 5.0; Middle: 5.68 - Control and Bottom; from left to right: 6.0,7.0,7.6 and 8.0

3.13 Effect of varying pH on radial growth and sporulation of BIO 56 T128. Top from left to right: 2.7,3.0,4.0, 5.0; Middle: 5.68 - Control and Bottom; from left to right: 6.0, 7.0, 7.6 and 8.0

3.14 Dual culture. Left to right: BIO T28, BIO T32, BIO T66 and BIO T128

3.15 Top: Bilayer plate - EGB 01 plated on BIO T28 (PDA + 0 . 0 2 0 ~ ~ - ' enl late^) Middle: Control I - EGB 01 plated only on PDA + 0.020 g ~ - ' enl late^) Bottom: Control 2 - EGB 01 plated only on PDA

3.16 Top: Bilayer plate - EGB 01 plated on BIO T28 (PDA + 0.01 ggL-' enl late^) Middle: Control I - EGB 01 plated only on PDA + 0.018 g ~ ' l enl late^) Bottom: Control 2 - EGB 01 plated only on PDA

3.17 Top: Bilayer plate - EGB 01 plated on BIO T28 (PDA + 0.016g~-' enl late') Middle: Control I - EGB 01 plated only on PDA + 0.016 g ~ - ' enl late^) Bottom: Control 2 - EGB 01 plated only on PDA

4.1 Rubber woodblock fully colonized by EGB 01 at 8 weeks of incubation

4.2 1 L of conidial suspension of the respective T. harzianum isolates, for the application as a soil drench at 1 Llseedling

4.3 Placement of artificial infection seedling in a pot filled with 73 113 of soil

4.4 An Illustration of the experimental layout of single (BIO 76 T32BIO T66), mixed (BIO T32 + BIO T66) and delayed treatments of infected plants using T. harzianum

4.5 The production of sporophores from control woodblocks 79 indicating viability of inocula

4.6 The disease classes describing the progression of disease signs 80 and symptoms.Top: Class 0 with healthy leaves and class 1 showing leaf necrosis, Middle: Class 2 with mycelia, Bottom: Class 3 with well developed sporophores and class 4 - dead

4.7 Trichoderma cfu treated soils observed as green colonies on 83 RBA

4.8 Disease Progression of EGB 01 on oil palm seedlings based 87 on disease severity index (DSI) between 0 to 24 w. a. i.

4.9 Mean of top dry weight of oil palm seedlings at 24 w. a. i.

4.10 Mean of root dry weight of oil palm seedlings at 24 w. a. i.

4.1 1 Seedlings of Control I (uninfected, untreated) uprooted at 24 w. a. i, showing good root volume (DSI=O)

4.12 Seedlings from Control I1 (infected, untreated) uprooted at 24 90 w. a. i, showing all seedlings succumbed to death with poor root development (DSI=86.67)

4.13 Seedlings from Treatment I (infected and treated with BIO T32) uprooted at 24 w. a. i, showing good root volume with few seedlings showing leaf chlorosis (DSF28.35)

4.14 Seedlings from Treatment I1 (infected and treated with BIO T66 alone) uprooted at 24 w. a. i, showing poor root development with more than 50% seedlings succumbed to death. I

4.15 Seedlings from Treatment I11 (infected and treated with mixture of BIO T32 & BIO T66) uprooted at 24 w. a. i, showing similar disease progression with Treatment I1 (DSI=76.67)

4.16 Seedlings from Treatment IV (infected and treated with BIO T32 at 6. w. a. i) uprooted at 24 w. a. i, showing good root mass with almost 50% of seedlings showing leaf chlorosis (DSI=45.0).

4.17 Mean reading of cfu/ g soil of T. harzianum at 5 cm depth between 0 to 22 w. a. i.

4.18 Mean reading of cfid g soil of T. harzianum at 15 cm depth between 0 to 22 w. a. i.

5.1 Experiment layout testing delivery system for T. harzianum (BIO T32) against G. boninense (EGB 01)

5.2 Illustration of experimental design for the effect of compost and surface mulch on growth of oil palm seedlings

5.3 Disease Progression of EGB 01 on oil palm seedlings based on disease severity index (DSI) over 24 w. a. i.

5.4 Mean reading of top dry weight of oil palm seedlings at 24 w. a. i.

5.5 Mean reading of root dry weight of oil palm seedlings at 24 w. a. i.

5.6 Mean reading of cfdg soil of T. harzianum for at 5 cm depth between 0 to 22 w. a. i.

5.7 Mean reading of cWg soil of T. harzianum for 15 cm depth between 0 to 22 w. a. i.

5.8 Top: Treatment I (ppf surface mulch); Oil palm seedlings infected with EGB 01, which was treated with conidial suspension of BIO T32 (mean of 1.68 x lo8 sporeslml) Bottom: Control I; Oil palm seedlings without infection and treatment.

5.9 Top: Treatment I (ppf surface mulch) - Oil palm seedlings infected with EGB 01, which was treated with conidial suspension of BIO T32 (mean of 1.68 x lo8 sporeslml). Bottom: Control I1 - Oil palm seedlings infected with EGB 01 without treatment.

5.10 Top: Control I - Oil palm seedlings without infection and treatment. Bottom: Treatment I1 (compost) - Oil palm seedlings infected with EGB 01, which was treated with conidial suspension of BIO T32 (1.67 x lo8 sporeslml)

5.11 Top: Treatment I1 (compost) - Oil palm seedlings infected with EGB 01, which was treated with conidial suspension of BIO T32 (1.67 x 10' sporeslml). Bottom: Control I1 - Oil palm seedlings infected with EGB 01 without treatment

5.12 Top: Control I - Oil palm seedlings without infection and treatment. Bottom: Treatment I11 (conidial suspension) - Oil palm seedlings infected with EGB 01 which was treated with conidial suspension of BIO T32 (1.68 x lo8 sporeslml)

5.13 Top: Control I1 - Oil palm seedlings infected with EGB 01 128 without treatment; Bottom; Treatment I11 (conidial suspension) - Seedlings infected with EGB 01 which was treated with only conidial suspension of BIO T32 (1.68 x lo8 sporeslml)

5.14 Top: Treatment IV (Hydrogel) - Oil palm seedlings infected with EGB 0 1 which was treated with hydrogel suspension of BIO T32 (1.67 x lo8 sporeslml). Bottom: Control I - Oil palm seedlings without infection and treatment.

5.15 Top: Treatment lV - Seedlings infected with EGB 01, which was treated with hydrogel suspension of BIO T32 (1.67 x lo8 sporeslml). Bottom: Control I1 - Oil palm seedlings infected with EGB 0 1 without treatment

5.16 Mean reading of top dry weight over 24 weeks

5.17 Mean reading of root dry weight over 24 weeks

5.18 Mean reading of leaf area measurement on 1 6th week of experiment

5.19 Mean reading of NPK percentage in oil palm seedlings

5.20 Comparison of seedlings of control (left) land seedlings treated 134 with compost (right)

5.21 Up rooted seedlings of compost (right) being compared with seedlings of control (left)

DSI

ANOVA

DMRT

PDA

MEA

TDW

RDW

PIRG

NPK

P P ~

LIST OF ABBREVIATIONS

: Disease Severity Index

: Analysis of Variance

: Duncan's Multiple Range Test

: Potato Dextrose Agar

: Malt Extract Agar

Top dry weight

Root dry weight

Percentage Inhibition of Radial Growth

Nitrogen, Phosphorus and Potassium

palm press fibre

colony forming unit

weeks after infection

gram

meter

: mililitres

liter

diameter

centimeter

kilogram

celcius

rotation per minute

CHAPTER I

INTRODUCTION

The oil palm, Elaeis guineensis, is the highest yielding among the oil-producing crops

(Ariffin et al., 2000). It is an important species in the tropical regions because of its two

main raw materials namely; palm oil and palm kernel oil. Palm oil commands an average

yield of about 4 tonnes oil ha-' year". In the year 2002, Malaysia produced 60% of the

world's palm oil with a total production of about 11 million tonnes (World Oils & Fats,

2002).

Like any other crop, the oil palm also faces a lot of pest and disease (P&D) tribulations.

From seed germination right up to field planting, the crop is exposed to several P&D

problems, some of which is caused by fungi. Some of the P&D problems faced by oil

palm industry are the basal stem rot, brown germ, upper stem rot, Rhinoceros beetles and

bagworm (Turner, 1981). Among these, the current most serious disease is Basal Stem

Rot (BSR). For the past 50 years or more, BSR had been causing serious damage to the

oil palm plantation in Malaysia. The disease is also prevalent in Indonesia, Zaire, Ghana,

Nigeria, Cameroon, San Tome, Principe, Angola, Rhodesia and Papua New Guinea

(PNG) (Turner, 1981) with incidence being relatively low in PNG (Pilotti, 2001).

The causal pathogen of this disease is the fungus Ganoderma. Not only does it attack oil

palms, it is also the causal agent of root and stem rots of other crops namely; coconut,